FIG. 4.
HrcN interacts with the conserved HrcL protein. (A) In vitro interaction studies with HrcL and HrcN. Strep-HrcN and Strep-HrcNG175C were immobilized on Sepharose matrix and incubated with an E. coli lysate containing HrcL-c-Myc. Bacterial total cell extracts (TE) and proteins eluted from the matrix (eluate) were analyzed by immunoblotting using c-Myc- and Strep epitope-specific antibodies. (B) Coimmunoprecipitation experiments with HrcN and HrcL in X. campestris pv. vesicatoria. Strains 85*ΔhrcL (ΔhrcL) and 85*ΔhrcN (ΔhrcN) carrying the empty vector (−) or synthesizing HrcL-c-Myc as indicated were incubated in the presence or absence of a c-Myc-specific antibody coupled to protein G-Sepharose as indicated. Bacterial total cell extracts (TE) and proteins bound to the Sepharose matrix (pull-down) were analyzed by immunoblotting using c-Myc- and HrcN-specific antibodies.